裙带菜配子体与孢子体的附生微生物群落组成分析

裙带菜(Undaria pinnatifida)是一种重要的大型褐藻,具有较高的经济和食用价值。藻类的附生微生物既能通过代谢产物调控宿主藻类的生长发育,特定条件下又可能导致病害。了解裙带菜附生微生物群落组成对研究裙带菜与附生微生物间的相互作用、种质资源的有效保存以及防治藻类病害等有重要意义。现有研究大多集中于海带和紫菜,关于裙带菜的附生微生物,特别是不同生活史的对比研究还很少。本研究通过16S rRNA基因高通量测序发现,裙带菜配子体和孢子体的附生细菌群落组成有明显差异,配子体样品中的细菌群落丰度和多样性均大于孢子体。配子体中变形菌门(Proteobacteria)(66.67%)为第一优势菌门,其次为拟杆菌门(Bacteroidetes)(13.48%)和蓝细菌门(Cyanobacteria)(11.13%),α-变形菌纲(Alphaproteobacteria)(34.58%)为第一优势菌纲,其次为γ-变形菌纲(Gammaproteobacteria)(31.01%);而孢子体中蓝细菌门(95.67%)占绝对优势,其次为放线菌门(Actinobacteria)(1.65%)和厚壁菌门(Firmicutes)(1.48%)。裙带菜样品经18S rRNA基因测序检测出链形植物(Streptophyta)、纤毛虫门(Intramacronucleata)、担子菌门(Basidiomycota)、顶复亚门(Apicomplexa)、节肢动物门(Arthropoda)、硅藻门(Bacillariophyta)、脊索动物门(Chordata)、腹毛动物门(Gastrotricha)、子囊菌门(Ascomycota)和毛霉菌门(Mucoromycota),其中,担子菌门、子囊菌门和毛霉菌门属于真菌,孢子体的真核微生物群落丰度大于配子体。本研究确定了裙带菜配子体和孢子体附生微生物群落组成以及不同细菌和真核微生物的相对丰度,结果表明,2个世代之间存在显著差异,为后续研究藻类宿主与微生物之间的相互作用、提高海带目褐藻种质保存技术提供了基础的数据支持。     

裙带菜孤雌生殖幼孢子体体细胞诱导2n♀配子体

以实验室内保存培养的裙带菜(Undaria pinnatifida)2n♀配子体为实验材料,经切碎培养,促进2n♀配子体成熟,排出卵进行孤雌生殖,形成幼抱子体.将0.5～1.0 cm的孤雌生殖幼孢子体的假根切除,从其体细胞中诱导出2n♀配子体.实验结果表明,在10～20℃范围内,培养20 d后,在15℃和20℃下的2n♀配子体排出的卵的孤雌生殖率分别为8.4%和7.4%,明显高于10℃下的1%0 20℃下诱导培养15d后就出现了2n♀配子体,培养60 d后,2n♀配子体的诱导率达到100%;15℃下培养30 d后才出现2n♀配子体,60 d后的诱导率为38%;而10℃下诱导培养45d后才开始出现2n♀配子体,60 d后的诱导率仅为12%.将诱导产生的2n♀配子体切碎,在20℃下培养10d后的成熟率为34%,60 d的成熟率达到100%.通过孤雌生殖幼孢子体诱导产生的2n♀配子体与正常♂配子体杂交形成了3n裙带菜幼孢子体.本研究发现了获得大量2n♀配子体的新方法,旨在为裙带菜三倍体幼苗规模化培育奠定基础.     

裙带菜3n、4n孢子体的人工育苗和海区栽培

使用室内保存培养的裙带菜2n♀♂配子体自交及与正常n♀♂配子体杂交,培育出裙带菜2n♀×n♂、n♀×2n♂两个组合3n幼孢子体和2n♀×2n♂组合的4n幼孢子体。经过40 d海区暂养后测定,3n幼孢子体两个组合平均长度分别为1.2 cm和1.1 cm,大于对照组合2n幼孢子体的0.9 cm和4n幼孢子体组合的0.7 cm。使用从短滨螺中提取的粗酶和R10纤维素酶的混合酶液将2n、3n幼孢子体酶解为原生质团,经Giemsa染色后观察到的染色体数分别为2n＝60,3n＝90。分苗后的海区栽培期间,在藻体的长度和重量方面,2n♀×n♂、n♀×2n♂两个3n孢子体组合间差异不显著(P>0.05),与对照组合2n孢子体之间的差异极显著(P<0.01)。在裙带菜进入繁殖期的5月上中旬,3n孢子体仍保持较快的生长速度,在5月20日测定两个组合的藻体长度和重量分别达到4.2 m、2.3 kg和4.0 m、2.2 kg,而对照组合2n孢子体仅达到2.3 m和1.3 kg。4n孢子体生长明显地慢于对照的2n孢子体(P<0.01),藻体生长最大值的时间为3月中旬,最大藻体长度和重量仅为1.6 m和1.1 kg。 试验结果表明,3n孢子体不发育,在繁殖期内无孢子叶形成,表现出不育的特性;4n孢子体形成孢子叶,但孢子叶很薄,孢子囊形成的数量少,发育水平也很低,经阴干刺激仅有极少量的孢子放出,显示出低育的特性。图2表1参15     

Projections of nutrient supply to a wakame (Undaria pinnatifida) seaweed farm on the Sanriku Coast of Japan

Wakame (Undaria pinnatifida) aquaculture is one of the major fishery industries on the Sanriku Coast of Japan. Low concentrations of nutrients in the autumn, when the germinated wakame sporophytes are installed at aquaculture facilities, induce defective growth, resulting in withering. To determine the characteristics of the nutrient variations in this region, we conducted hydrographic measurements off the coast of Iwate Prefecture. The measured results showed that in the autumn, vertical mixing pumped nutrients from the deeper layers to the mixed layer, where nutrients had been depleted during the summer. Based on these results, we developed a one‐dimensional model incorporating vertical mixing by atmospheric cooling to project the vertical distribution of the nutrient concentrations. By conducting ensemble calculations, we created a probability projection calendar for the nutrient supply up to 50 days ahead. This information was provided to wakame farmers before they installed the germinated sporophytes. The projected probability calculated from the initial conditions on 11 October 2016 was 0% until 25 October and gradually increased to 85% on 30 November. The nitrate concentration detected by an ultraviolet nitrate sensor moored at a wakame farm revealed that the nutrient concentrations reached a critical threshold on 18 November. The projected probability on that date was 75%; therefore, the projection was successful. The observed temporal evolution of the temperature, salinity, and nutrient concentrations was well reproduced by our model. The information obtained using this model is expected to reduce the risk of wakame withering and improve the efficiency of wakame production.     

桑沟湾养殖海带生长的模型预测

海带(Saccharinajaponica)是一种常见的大型经济褐藻,是海洋生态系统重要的初级生产者,也是我国北方沿海主要的养殖藻类。本研究旨在建立海带个体生长数值模型,并以此预测中国北方近海大规模筏式养殖条件下海带的生长情况。本研究以桑沟湾养殖海带为例,利用可视化模型软件STELLA描述海带生长的关键过程及其与环境参数的关系,以净生长量(N_(growth))=总生长量(G_(growth))–呼吸作用(resp)–枯烂(E_(kelp))为基本框架,模拟和预测海带的生物量和叶片长度变化。海带的总生长用光照、温度、盐度、海带体内营养盐(包括N和P)等强制函数定义,其中,光照参数来自桑沟湾气象记录,盐度、温度和营养盐为现场调查实测值。模型模拟桑沟湾养殖海带的长度与干重结果与实测值的拟合度R~2值分别为0.936、0.963,说明该模型能够很好反映海带的真实生长情况。可靠的个体生长模型是评估海带养殖容量的基础,并可为水产养殖区的空间规划提供决策依据。     

Distribution patterns of five pleuronectid species on the continental slope off the Pacific coast of northern Honshu,Japan

Information on the distributions of demersal and benthic fishes is fundamental for stock assessment and management. Spatiotemporal changes in the distribution patterns of five pleuronectid species (flathead flounder Hippoglossoides dubius, Kamchatka flounder Atheresthes evermanni, roughscale sole Clidoderma asperrimum, slime flounder Microstomus achne, and Korean flounder Glyptocephalus stelleri) off the Pacific coast of northern Honshu, Japan, at depths of 150–900 m, were examined using a generalized linear mixed model (GLMM). Densities of flathead and Korean flounder were highest in the southernmost area, where the fish were small. The body lengths of both of these species increased from 2003 to 2008, suggesting that an abundant year class was recruited in 2003. The density of Kamchatka flounder was highest in the northern area. In roughscale sole and slime flounder, there were no distinctive annual and latitudinal trends in the density distributions. The density distribution of Korean flounder was bimodal; the peaks were at depths of 210 and 410 m. The body length increased as the depth increased from 150 to 410 m, and then decreased from 410 to 550 m. Moreover, “bigger–deeper” trends were observed in flathead, Kamchatka and slime flounder.     

裙带菜优质高产相关因素的探讨

对裙带菜产业长期存在的一些生产、管理上的弊端作了探索,就裙带菜半人工苗和全人工苗的配置、海域布局、合理放养密度、科学采收以及生产考核制度等方面提出了一些切合当前生产实践的积极措施。     

Effects of a highly purified fucoidan from Undaria pinnatifida on growth performance and intestine health status of gibel carp Carassius auratus gibelio

A six‐week feeding trial was conducted to determine the effects of different concentrations of fucoidan (1 g/kg, 10 g/kg and 30 g/kg; w/w) from Undaria pinnatifida on gibel carp (Carassius auratus gibelio). Our results demonstrated that 30 g/kg fucoidan significantly increased (p < .05) growth performance, intestinal digestive enzyme activities, acid phosphatase activity and immunoglobulin M content. Histological examinations revealed that gibel carp receiving 30 g/kg fucoidan had significant higher abundance of mucin‐containing goblet cells in middle and distal intestine as compared with control treatment (p < .05). Intestinal microbiota analysis showed that 30 g/kg fucoidan supplementation significantly increased (p < .05) the abundance of Cetobacterium and Aeromonas, but lowered (p < .05) the prevalence of pathogenic bacteria Plesiomonas and a mucin‐degrading bacterium Mucinivorans. Furthermore, RNA‐seq and RT‐qPCR analysis indicated that 30 g/kg fucoidan caused significant changes (p < .05) in the expression of genes involved in immune regulation (such as interleukin‐8 and cyclooxygenase), signal transduction (such as phosphatidylinositol‐4,5‐bisphosphate 3‐kinase and protein kinase B) and nutrition utilization (maltase–glucoamylase and muscarinic acetylcholine receptor 3). Together, the current study shows that fucoidan supplementation could elevate the activity of intestinal digestive enzymes, modulate intestinal microbial communities and potentiate a higher state of immune readiness, which might consequently improve growth performance and intestine health status of gibel carp.     

Comparison of environmental conditions in two representative oyster farming areas: Hiroshima Bay,western Japan and Oginoham a Bay (a branch of Ishinomaki Bay), northern Japan

ABSTRACT:   Sea water environmental conditions over annual cycles were investigated and compared between two oyster farming areas, western Hiroshima Bay and Oginohama Bay (a branch of Ishinomaki Bay) in Miyagi Prefecture, to appropriately manage oyster culture or more efficiently utilize farming areas. The environmental parameters of temperature, salinity, nutrient concentrations (NO₂–N, NO₃–N, NH₄–N, PO₄–P, and SiO₂–Si) and size-fractionated chlorophyll- a (<0.2, 2–20, >20µm), and abundances of microzooplankton were measured in each bay at the surface, and 2 and 5 m depth layers. Differences in the annual mean values and results with monthly paired Student's t -tests showed that salinity was lower, and temperature, nutrient (especially PO₄–P) and chlorophyll- a concentrations, and abundance of microzooplankton, were higher in Hiroshima Bay than in Oginohama Bay. Differences in environmental conditions between inshore and offshore areas of each bay suggest that inflows of river water in western Hiroshima Bay and sea water from offshore had the most significant effects on the environmental conditions. It is concluded that such oceanographic and biological differences strongly affect the oyster farming system, especially regarding the optimum usage of offshore areas in Summer under clean, cold and stable seawater conditions, rather than food quantity in Hiroshima Bay, and under more abundant food conditions in Oginohama Bay.     

Epiphytism on Blades of the Edible Kelps Undaria pinnatifida and Saccharina latissima Farmed under Different Abiotic Conditions

The kelps Undaria pinnatifida and Saccharina latissima are economically valuable edible seaweeds. Although epiphytism is a major problem affecting the productivity and quality of farmed seaweeds, studies of its effects on kelp aquaculture are scarce. We investigated the effects of epiphytism in two U. pinnatifida and S. latissima culture grounds located in two different parts (exposed and sheltered) of a bay (ría) in Galicia (NW Spain). Epiphytic organisms on blades were identified and their abundance was documented. The percentage of the total blade covered with epiphytes was measured for both kelp species in order to estimate the blade biomass that must be discarded during processing for human consumption. Algae, hydroids, and crustaceans were the most common epiphytic organism on kelp blades; colonies of the hydroid Obelia geniculata and harpacticoid copepods were the most abundant epiphytes. Abundance of epiphytes on the blades of both kelps was significantly higher for fronds growing in the sheltered area of the bay compared to those farmed at the exposed location. Different environmental conditions and other factors, such as the proximity of mussel cultures to kelp cultures, may be important contributors to epiphytism. The results presented here can be used to devise better ways for kelp aquaculture facilities to diminish the negative effects of epiphytism.     

Effect of temperature, age class, and growth on induction of aestivation in Japanese sandeel (Ammodytes personatus) in Ise Bay, central Japan

We investigated the effects of temperature, age class, and growth on induction of aestivation in the Japanese sandeel (Ammodytes personatus) by means of rearing experiments and field surveys in Ise Bay, central Japan. Acoustic surveys and sampling of shoaling and aestivating sandeels indicated that the sandeels completed aestivation when the water temperature increased to 17–20°C. A rearing experiment showed that the mortality of sandeels reared without sand was greater than it was with sand, under replete feeding conditions. In a non‐aestivation rearing experiment without sand, the condition factor of the sandeels decreased when water temperatures exceeded 20°C, although they were fed to repletion. These results suggest that aestivation is induced by rising seawater temperature and plays an important role in energy conservation. The age‐1 group tended to start aestivation earlier than the age‐0 group in the field. Within the age‐0 group, larger sandeels tended to enter aestivation earlier. These observations indicate that fat reserves are an important cue to start aestivation.     

南美白对虾的性腺发育、交配、交卵和受精

以池养南美白对虾（Penaeus vannamei）亲虾进行人工促熟，并使之交尾、产卵、受精。实验在上海金山区抗州湾河口低盐地区一套自行设计的水循环系统中进行，育苗用水通过兑配使盐度为27-28。育苗期间基本不换水和移动亲虾。亲虾205尾，雌雄比约1：1。雌虾切除眼柄后3-4d，性腺即开始发育，6-7d产卵，但未受精。手术后约30d，亲虾出现追尾现象、随之发生交配并出现受精卵。实验进行了约60d，共收集虾卵456.7万粒，其中受精卵出现后所产卵为137万粒，通过设置在水循环系统内的集卵网袋，可收集到95％以上的卵，共获受精卵61万粒，平均日受精率为44.5％，无节幼体25.1万尾，受精卵的平均孵化率为41.2％，最高达87.5％。文中还重点讨论了雌雄亲虾性腺发育的特点，产卵交配规律，以及亲虾规格大小、饵料种类、水质状况对南美白对虾交配和受精的影响。     

南美白对虾的性腺发育、交配、产卵和受精

对池养南美白对虾 (Penaeusvannamei)亲虾进行人工促熟 ,并使之交尾、产卵、受精。实验在上海金山区杭州湾河口低盐地区一套自行设计的水循环系统中进行 ,育苗用水通过兑配使盐度为 2 7～ 2 8。育苗期间基本不换水和移动亲虾。亲虾 2 0 5尾 ,雌雄比约 1∶1。雌虾切除眼柄后 3～ 4d ,性腺即开始发育 ,6～ 7d产卵 ,但未受精。手术后约 30d ,亲虾出现追尾现象、随之发生交配并出现受精卵。实验进行了约 6 0d ,共收集虾卵 4 5 6 .7万粒 ,其中受精卵出现后所产卵为 137万粒 ,通过设置在水循环系统内的集卵网袋 ,可收集到 95 %以上的卵 ,共获受精卵 6 1万粒 ,平均日受精率为 4 4 .5 % ,无节幼体 2 5 .1万尾 ,受精卵的平均孵化率为 4 1.2 % ,最高达 87.5 %。文中还重点讨论了雌雄亲虾性腺发育的特点 ,产卵交配规律 ,以及亲虾规格大小、饵料种类、水质状况对南美白对虾交配和受精的影响     

Effects of hypoxia on principal prey and growth of flathead flounder Hippoglossoides dubius in Funka Bay, Japan

ABSTRACT:   Effects of hypoxia on the principal prey and growth of flathead flounder Hippoglossoides dubius were studied in Funka Bay. Of the three dominant year-classes that occurred in recent years, the 1995 year-class was small in total length at age ≥ 3 and low in condition factor at age ≥ 2. Ophiuroids (almost Ophiura sarsi ), which were the dominant prey in the 1980s, were of little importance and instead, crustaceans such as mysids, natant decapods and pelagic amphipods, bivalves and fish were important prey items for H. dubius in 2000–2001. In addition, the feeding intensity of H. dubius in 2000–2001 was lower than that in the 1980s. These facts are closely related to a reduction of prey abundance, particularly ophiuroids. It seems that the hypoxia that occurred in the central part of the bay during the summer and autumn of 1995–1997 caused the poor food supply and low growth rate at ages 2–4 of the 1995 year-class.     

Improved growth of flathead flounder Hippoglossoides dubius in hypoxic waters in Funka Bay,Japan

To identify the factors that influence the growth rate of flathead flounder Hippoglossoides dubius in Funka Bay, Japan, we studied temporal changes in the growth of this species and its prey from 1989 to 2012. The lowest growth rate of H. dubius was recorded in the 1995 year-class, which had experienced severe hypoxia between 1995 and 1997. However, the highest growth rate was recorded in the 2007 year-class, which had experienced hypoxia between 2009 and 2011. In 2000 and 2001, small-sized flounder (<200 mm TL) from the 1995 year-class were feeding on mysidaceans, whereas those from other year-classes fed on small Ophiura spp. (<9 mm in disc diameter) in 2009, 2011, and 2012. Small Ophiura spp. were more abundant in 2007 and 2011 than in 2002 and 2003. Successful recruitment of Ophiura spp. took place between 2007 and 2011 and cohorts from these years could tolerate hypoxia. This suggests that food availability improved under hypoxic conditions, which led to improved growth in small flounders. Large flounders (≥200 mm TL) from all year-classes fed on prickleback fishes (Stichaeidae) and walleye pollock Gadus chalcogrammus juveniles. The growth of large flounders was enhanced by an increase in the abundance of Stichaeidae fishes and G. chalcogrammus juveniles.